LOCUS       sequence-413258-        4122 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Riboprobe generation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4122)
  AUTHORS   Park SJ, Silic MR, Staab PL, Chen J, Zackschewski EL, Zhang G
  TITLE     Evolution of two-pore domain potassium channels and their gene 
            expression in zebrafish embryos.
  JOURNAL   Dev Dyn. 2024 Jan 25. doi: 10.1002/dvdy.690.
  PUBMED    38270285
REFERENCE   2  (bases 1 to 4122)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Dev Dyn. 
            2024 Jan 25. doi: 10.1002/dvdy.690."
COMMENT     Sequence Label: pENTR-D-Topo-kcnk5a
FEATURES             Location/Qualifiers
     source          1..4122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     6..28
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     20..37
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     21..37
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    53..152
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1731..1830)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1847..1866)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1848..1866)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1871..1887)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1871..1887)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1989..2008)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2000..2809
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2067..2086)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2956..3544
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3445..3464
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3698..3715
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3874..3901)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3993..4079)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gttttcccag tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc cccaaataat
       61 gattttattt tgactgatag tgacctgttc gttgcaacaa attgatgagc aatgcttttt
      121 tataatgcca actttgtaca aaaaagcagg ctccgcggcc gcccccttca ccatggtgga
      181 caagggacct cttctgactt ctgctatcat tttctacctg tcgattgggg cagcaatatt
      241 ccaagtcatc gaagaaccga actgggagat cgccgtgagg aaatatagag cagacaagga
      301 gagcatattg aaacaatacc cctgccttac caaggctgac ctggactaca ttctagaggt
      361 ggtgtccgat gctgcaggac aaggcatcac catcacaggc gacaaaacct ttaacaactg
      421 gaactggccc aatgccgtca tctttgcagc cacagtgatc acaactatag gttatggaaa
      481 tattgctcca aaaactcctt cagggagggt tttctgtata ttttatggtc tctttggagt
      541 gccactgtgt ttcacttgga tcagtgaact tggaaagttt tttggtggaa gagctaaaca
      601 cctcggatgg tatcttacaa agaaaggagt aacattgcgg aaaacgcagc tcacctgtac
      661 agctgtcttt ctgttatggg gtgtgctgat ccatttggtg attcctccat ttgttttcat
      721 gactcaagaa ggctggacct atatcgaggg cttgtatttt tctttcgtca ctttgacaac
      781 tattggtttt ggtgatcttg tagcaggtgt tgacccaaat gctgaatatc ctactctcta
      841 tcgctacttt gttgaggtgt ggatttatct tggtctggca tggctgtctt tgtttttcaa
      901 ttggaaggtg cgtatggttg tggaggctca caaagctctg aagaagcacc gtaaaaggcg
      961 taggctttca ctggatgagt tacagctcaa ggaaagtcat aagacactac accttccccc
     1021 aactcccaat gatgtcaaca tcttcagctt cctgtccaag aaaaaggaag gttacaatga
     1081 cttaattaag caaataggtg ctgacaaagg agaaaatcag tgtgtcagca ctatgaatag
     1141 tagaaagaac gcagccgcaa agcgctcaaa aagctgcagc gatgctctct ctataaatgg
     1201 aaatactatt gttagtctag acggctcacc tcgactgaag cgccattata gcttcagtga
     1261 tagagttgct gtagcactct caaaatccaa gggctacctt tttagccaag aggaatgtct
     1321 tctcatgact gagaagctag aggaaggaga tactgacttg cagtgcatgg gaatgtctgg
     1381 gaatcagttg gacaaggagg caggagagag ccatcaagct cttatatatg aaggctgcag
     1441 aaatggtccg atcacatggg actccaaagg tcaccagacc aacatgtttc ctaatgctaa
     1501 cataaccttc attgatgagg ataattttgt aaacaataac cgggaggatg aaaatgactt
     1561 tatgcctagc ttcaaatcag atgagccaga tttcagaaag gaagaggact cagaatctga
     1621 aacctctgta ttcaccagat gtggctcaga ccatggacaa tcctacgagc ggctggtgga
     1681 agagtattcc aaagaggaca acactgacat ttaaaagggt gggcgcgccg acccagcttt
     1741 cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt
     1801 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
     1861 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     1921 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     1981 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     2041 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     2101 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     2161 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     2221 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     2281 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     2341 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     2401 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     2461 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     2521 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     2581 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     2641 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     2701 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     2761 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     2821 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     2881 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     2941 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3001 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3061 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3121 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3181 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3241 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3301 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3361 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3421 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3481 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3541 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3601 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     3661 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     3721 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     3781 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg
     3841 taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc
     3901 tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt
     3961 tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc
     4021 gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga
     4081 tgcctggcag ttccctactc tcgcgttaac gctagcatgg at
//
